Njambere E N, Clarke B B, Bonos S A, Murphy J A, Buckley R, Tirpak S, Zhang N
Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901.
Plant Dis. 2011 Jan;95(1):78. doi: 10.1094/PDIS-08-10-0606.
Waitea circinata var. circinata was first reported as the causal agent of brown ring patch on annual bluegrass (Poa annua L.) in the United States in 2007 (2). In early April to mid-June of 2009, circular to irregularly shaped yellow rings resembling symptoms of this disease were observed on an annual bluegrass putting green at Rutgers University in North Brunswick, NJ. Severely infected foliage eventually turned brown as the disease progressed. During the same time period, similar disease symptoms were observed on creeping bentgrass (Agrostis stolonifera L.) from a golf course in Bedminster Township, NJ. The disease reappeared in both locations in April of 2010. Five additional samples with similar symptoms on creeping bentgrass and annual bluegrass were received at Rutgers Diagnostic Laboratory from Paramus, Madison, Allamuchy, and Farmingdale, NJ between late April and early May of 2010. Portions of diseased leaf and sheath tissue that displayed symptoms of the disease were disinfested for 1 min in 0.5% NaOCl, rinsed with sterile distilled water, and plated on potato dextrose agar (PDA) amended with 50 mg/liter of streptomycin sulfate. At the first sign of fungal growth, single hyphal tips were transferred to PDA. After 1 week at 25°C, white-to-orange mycelial colonies formed in culture and eventually turned brown with age. Minute sclerotia (≤3 mm), which followed the same color development pattern, formed within 10 days. These features are consistent with those described of W. circinata var. circinata (2,3). The internal transcribed spacer (ITS) region of the ribosomal RNA gene was amplified using primer pair ITS1/ITS4 and sequenced with ITS4 (GenBank Accession Nos. HQ166065 to HQ166071). BLASTn analysis of the ITS sequences showed a 99 to 100% similarity to W. circinata var. circinata sequences deposited in GenBank (1,2). Pathogenicity tests were conducted in 2010 using 6-week-old creeping bentgrass seedlings cv. Declaration inoculated with colonized oat grain that had been autoclaved and then infested with the Bedminster Township isolate. Eight colonized oat grains were uniformly spread around the crowns of seedlings grown in 10-cm-diameter pots. Control plants were treated with autoclaved grain. Plants were incubated at 25°C and high humidity maintained by misting the plants three times per day. Within 3 days postinoculation, foliage near infested grain turned chlorotic. All foliage in pots became completely blighted and spherical orange-brown sclerotia were observed on leaf sheaths by the eighth day. W. circinata var. circinata was consistently reisolated from inoculated plants (as confirmed by isolate morphology and ITS sequencing) but not from control plants. The ITS sequence data, morphological characters of the isolates, and pathogenicity tests demonstrate that W. circinata var. circinata is present in New Jersey. To our knowledge, this is the first report of W. circinata var. circinata infecting turfgrass in New Jersey. References: (1) C. M. Chen et al. Plant Dis. 93:906, 2009. (2) K. A. de la Cerda et al. Plant Dis. 91:791, 2007. (3) T. Toda et al. Plant Dis. 89:536, 2005.
2007年,在美国,环绕丝核菌圆斑变种首次被报道为一年生早熟禾(Poa annua L.)上褐色环斑病的致病因子(2)。2009年4月初至6月中旬,在新泽西州北不伦瑞克市罗格斯大学的一个一年生早熟禾果岭上,观察到了圆形至不规则形状的黄色环,类似于这种病害的症状。随着病情发展,严重感染的叶片最终变成褐色。在同一时期,在新泽西州贝德明斯特镇一个高尔夫球场的匍匐翦股颖(Agrostis stolonifera L.)上也观察到了类似的病害症状。2010年4月,这两个地方的病害再次出现。2010年4月下旬至5月初,罗格斯诊断实验室从新泽西州帕拉默斯、麦迪逊、阿拉穆奇和法明代尔收到了另外5份在匍匐翦股颖和一年生早熟禾上有类似症状的样本。将表现出病害症状的病叶和叶鞘组织部分在0.5%次氯酸钠中消毒1分钟,用无菌蒸馏水冲洗,然后接种到添加了50毫克/升硫酸链霉素的马铃薯葡萄糖琼脂(PDA)上。在真菌生长的最初迹象出现时,将单个菌丝尖端转移到PDA上。在25°C下培养1周后,培养物中形成了白色至橙色的菌丝菌落,随着时间的推移最终变成褐色。在10天内形成了微小菌核(≤3毫米),其颜色变化模式相同。这些特征与已描述的环绕丝核菌圆斑变种的特征一致(2,3)。使用引物对ITS1/ITS4扩增核糖体RNA基因的内部转录间隔区(ITS),并用ITS4进行测序(GenBank登录号:HQ166065至HQ166071)。对ITS序列的BLASTn分析表明,其与GenBank中 deposited的环绕丝核菌圆斑变种序列有99%至100%的相似性(1,2)。2010年进行了致病性测试,使用6周龄的匍匐翦股颖品种Declaration的幼苗,接种经过高压灭菌然后接种了贝德明斯特镇分离株的定殖燕麦粒。8个定殖燕麦粒均匀地散布在直径10厘米花盆中生长的幼苗的冠部周围。对照植物用高压灭菌的谷物处理。将植物在25°C下培养,并通过每天对植物喷雾3次来保持高湿度。接种后3天内,受侵染谷物附近的叶片变黄。到第8天,花盆中的所有叶片都完全枯萎,在叶鞘上观察到球形橙褐色菌核。始终能从接种的植物中重新分离出环绕丝核菌圆斑变种(通过分离株形态和ITS测序确认),但从对照植物中未分离到。ITS序列数据、分离株的形态特征和致病性测试表明,环绕丝核菌圆斑变种存在于新泽西州。据我们所知,这是环绕丝核菌圆斑变种感染新泽西州草坪草的首次报道。参考文献:(1)C.M. Chen等人,《植物病害》93:906,2009年。(2)K.A. de la Cerda等人,《植物病害》91:791,2007年。(3)T. Toda等人,《植物病害》89:536,2005年。